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Luminex® Assays
High-throughput Multiplex Bead Based Assays
Luminex assays are based on xMAP® technology (multi-analyte profiling beads) enabling the detection and quantitation of multiple RNA or protein targets simultaneously. The xMAP system combines a flow cytometer, fluorescent-dyed microspheres (beads), lasers and digital signal processing to efficiently allow multiplexing of up to 100 unique assays within a single sample.
Panomics has an aggressive program for the development of a broad range of assays for the Luminex platform and similar instruments based on the xMAP technology. Currently we have hundreds of RNA targets available in 3–30 plex assays using our using our QuantiGene Plex Reagent Systems, with new targets being added weekly. Our Procarta® Human, Mouse and Rat Assays can quantitatively measure cytokines and chemokines from a variety of sample sources, including serum and plasma. We’ve recently expanded our family of assays designed to measure protein expression and monitor protein modifi-cations/activation states in diverse matrices.
We are also proud to be one of the few Luminex Certified Developers. Luminex have recognized us as having both unique and extensive assay development capabilities. We are happy to discuss your specific needs and develop and validate your assay to the same exacting standards as our commercially available assays.
Luminex Assays from Panomics
Gene Expression—quantitatively measure up to 30 different RNA transcripts
Transcription Factor—Profile up to 40 different active TFs from a single sample
Cytokine/Chemokines—quanti-tatively measure up to 33 different secreted proteins in serum, plasma or cell culture supernatant in human and mouse samples
SH2 Domains—Profile phospho-tyrosine interactions with 30 SH2 protein binding domains
Custom Built Assays—If you can’t find what you’re looking for, we can custom design and build your assay. As one of the few Luminex Certified Developers, you can be assured of quality and a speedy response.
CE
mRNA
LE
BL
CP
CaptureBead
2.0 Pre-Ampli�er
2.0 Ampli�er withbiotinylated Label Probe
Streptavidin Phycoerythrin
a)
b)
Dried Blood Spots
Whole Blood orPAXgene Blood
FFPE Sections
Animal Tissues
Cultured Cells
Step 1: Release Target RNA
Cells are lysed to release RNA.
Step 3: Signal Ampli�cation
a) Sequential hybridization of the2.0 Pre-Ampli�er, 2.0 Ampli�erand biotinylated Label Probe, respectively, for an hour at 50°C.
b) Binding with Streptavidin-conjugated Phycoerythrin (SAPE) at room temperature for 30 minutes.
Step 4: Detection
The sample is analyzed on a Luminex* instrument. The level of SAPE �uorescence is proportional to the amount of mRNA transcripts captured by the bead.
* Bio-Plex suspension array system or other Luminex-based array systems.
Step 2: Target RNA Capture
Speci�c mRNA transcripts are captured to their respective beads through a Capture Extender (CE) Capture Probe (CP) interaction during an overnight hybridization at 54°C.
QuantiGene Plex 2.0—Taking Multiplexed Gene Expression to the Next Level
QuantiGene Plex 2.0 and Luminex
QuantiGene® Plex offers high reproducibility and ease-of-use that make it the perfect assay to bridge the technology gap when studying many genes in a limited number of samples and study-ing a few genes in a large number of samples. With QuantiGene Plex, researchers can easily perform multiplexed analyses from rare or volume-limited samples and can compare results across differ-ent samples, experiments and laboratories. Profiling many genes simultaneously in a single reaction directly from cultured cell or whole blood lysates, or fresh, frozen or FFPE tissue homogenates, can be accomplished without the need for RNA purification, reverse transcription, or amplification.
QuantiGene Plex 2.0 assays combine branched DNA (bDNA) signal amplification technology and xMAP (multi-analyte profiling) beads to enable simultaneous quantification of multiple RNA targets directly from cultured cell or whole blood lysates; fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tissue homogenates; or purified RNA preparations. Clinically proven Branched DNA technology is a sandwich nucleic acid hybridization assay that provides a unique approach for RNA detection and quantification by amplifying the reporter signal rather than the target sequence. By measuring the RNA at the sample source, the assay avoids variations or errors inherent to extraction and amplification of target sequences.
Assay Specifications
Limit of Detection ≤ 2,000 transcripts/assay well
Limit of Quantitation ≤ 5,000 transcripts/assay well
Linear Dynamic Range ≥ 3 logs
Assay CV ≤ 15% intra-assay; ≤ 20% inter-assay
Compatible Sample Types
Cultured cells, whole blood, PAXgene blood or dried blood spots, fresh/frozen tissues, FFPE samples, purified RNA
Assay Format 96-well plate
Targets/well 3–30
QuantiGene Plex Applications
• Prospective/retrospective studies using whole blood or FFPE samples
• Biomarker validation
• Predictive toxicology
• Microarray validation
• Secondary screening
• RNAi knockdowns and monitoring of “off-target” effects
Demonstrated Performance with Clinically Relevant Sample Types
Using QuantiGene Plex 2.0 in High Throughput Applications
Many potential drugs that specifically target a particular protein considered to underlie a given disease have been found to be less effective than hoped, or to cause significant side effects. The intrinsic robustness of living systems against various perturbations is a key factor that prevents such compounds from being successful.
By including screening measurements in a more integrated manner using chemical genomic approaches, i.e. associated pathway elements, dose response, “off target” targets, the likelihood of identifying more robust compounds (SME’s) or biologicals that have a higher chance of ulti-mate success will increase significantly. At the same time promising candidates that would have ultimately failed at a later stage in the development process will be identified during screening, enabling higher attrition rates supporting the ultimate goal of “failing faster”.
QuantiGene Plex 2.0 is ideally suited to deliver cost effective multiplex data for these new high value contextually relevant assays. Benefits to consider:
• Target additional pathway elements not just primary genes of interest
• Get earlier indication of toxicology profiles and stress indicators
• Develop dose response profiles
• 384 well or 96 well plate formats are supported
• Assay can be readily automated
• Our patent pending plex/plex methodology reduces cost and labor considerably
• Compatible with the Luminex HTS system
0.1
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10000
0.1 1 10 100 1000 10000
QuantiGene Plex
Qua
ntiG
ene
Plex
2.0
Housekeeping RNAs
Cytokine RNAs
Values in the graph demonstrate the expected induction of 4 cytokine RNAs in 10 µL of whole blood samples treated with LPS. No significant changes were detected in the expression of 8 housekeeping RNAs. Data from QuantiGene Plex 1.0 and QuantiGene Plex 2.0 assays had a correlation coefficient of 0.9995.
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PDH
RPLP
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PGK1
HPR
T1
PPIB
POLR
2A
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RPL1
9
RPL3
2
RPS3 IL-8
Housekeeping RNAsTarget
Fold
-cha
nge
(+PM
A/-
PMA
) QuantiGene Plex
QuantiGene Plex 2.0Values in the graph demonstrate the expected induction of IL-8 in FFPE preparations of HeLa cell pellets treated with PMA. No significant changes were detected in the expression of 10 housekeeping RNAs. Data from the QuantiGene Plex 1.0 and QuantiGene Plex 2.0 assays had a correlation coefficient of 0.9999.
QuantiGene Plex Publications
1. Gupta, A., et al., Role of protein C in renal dysfunction after polymicrobial sepsis. J Am Soc Nephrol, 2007. 18(3): p. 860-7.
2. Flagella, M., et al., A multiplex branched DNA assay for parallel quantitative gene expression profiling. Anal Biochem, 2006. 352(1): p. 50-60.
3. Zheng, Z., Y. Luo, and G.K. McMaster, Sensitive and quantitative measurement of gene expression directly from a small amount of whole blood. Clin Chem, 2006. 52(7): p. 1294-302.
4. Zhang, A., et al., Small interfering RNA and gene expression analysis using a multi-plex branched DNA assay without RNA purification. J Biomol Screen, 2005. 10(6): p. 549-56.
Assay Highlights
Quantitatively measure multiple RNA targets simultaneously with unparalleled accuracy and precision
RNA quantitation directly from cultured cells, whole blood, or fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tissue
– No RNA purification – No reverse transcription – No target amplification
Simple Assay Workflow
Widely used in biomarker validation, microarray validation, predictive toxicology and secondary screening
Procarta Transcription Factor Plex—Luminex Assays for Transcription Factor Profiling
About Transcription Factors
Transcription Factors (TFs) are highly conserved proteins that bind to DNA and initiate transcription of a given gene. A single extracellular stimulus can trigger multiple signaling pathways, and these in turn can activate multiple TFs to mediate the inducible expression of target genes.
The Procarta TF Plex assay is a profiling assay for monitoring the activation of TFs. We have developed two panels (40-plex and 43-plex) which are Luminex based to profile and measure activated TFs. The 96-well plate format enables high-throughput profiling of the DNA binding activity of TFs in multiple samples with high sensitivity.
Key Applications
• Profile the activities of multiple TFs upon a given drug stimulus
• Monitor off-target effects upon a given drug treatment
• Confirm cell signaling pathways using the TF Plex Assay
How It Works
Our novel, Procarta TF assay allows for the profiling of multiple TFs from a variety of sample types including cell lysates and nuclear extracts. Up to 40 TFs can be analyzed in one well.
bead1
bead2
Cis2Cis1
Cis1
Cisn Cis2
Cisn
bead3
Step 3: Denature cis elements by heat and aneal to Luminex beads with anti-sense sequence
Step 4: Add streptavidin PE to sample and read on Luminex machine
Cis1
Cisn
Cisn
Cis2
Cis2TF2
TFn
TF1
Cis1
Cis1
Cisn
Cis2
TF2
TFn
TF1
Step 1: Incubation of thecis element probes with the nuclear extract or whole cell lysate in 96 well plate
Step 2: Transfer probeTF mixture to separation plate and wash TF bound probe mix. Denature cis elements away from TFs
Procarta Transcription Factor Plex—Luminex Assays for Transcription Factor Profiling
TF 40-plex assay: Nuclear Extract from HeLa +/- PMA
MFI
0
2000
4000
6000
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12000
14000
Nuclear Extract –PMANuclear Extract +PMA
RUNX
/AML AP
1
AP2 AR
ATF2
NF-Y
CEBP
FAST
1
C-MYB
CREB
E2F1
-1
ELK1 ER
ETS/
PEA
FKHR
-1
GATA
-1
GR/P
R
HIF-1
HNF1
IRF1
ISRE
MEF-2
MYOD NF-1
NFAT
NF-E
1 (YY
1)
NF-E
2
NFKB
1
OCT
P53
PPAR
PAX3
SMAD
STAT
1
STAT
3
STAT
4
STAT
5
NKX-
2.5
BRN3
PAX5
Probe only
Procarta TF Plex Assay Confirmed by EMSA
The activity of 40 different TFs were profiled using the Procarta Transcription Factor Plex Assay. Nuclear extracts were prepared from serum starved HeLa cells subse-quently stimulated with PMA or a vehicle control for 4 hours. Extracts were used on the Procarta TF Plex assay and the EMSA Gel Shift assays.
1 2 3
AP-1
1 2 3
NF-E2
1 2 3
STAT4
1 2 3
NF-1
Create your own TF Plex Panel
You have the option to either order the full 40 plex or choose TFs from either panel to create your own unique 3-39 plex.
RUNX/AML ELK-1 ISRE OCT
AP-1 ER MEF-2 p53
AP-2 ETS/PEA MYOD PAX-3
AR FAST-1 NF-1 PAX-5
ATF-2 FKHR-1 NFAT PPAR
BRN-3 GATA-1 NF-E1/YY1 SMAD
CEBP GR/PR NF-E2 STAT-1
C-MYB HIF-1 NFkB STAT-3
CREB HNF1 NKX-2.5 STAT-4
E2F-1 IRF-1 NF-Y STAT-5
Procarta TF Panel 1 (40 TFs)
Visit our website to see the most current list.
Procarta TF Panel 2 (43 TFs)
Create your own TF Plex Panel
You have the option to either order the full plex sets from Panel 1 or Panel 2 or choose TFs from within either panel to create your own plex set.
ALF-1/TAL-1 ELF-1 KPF-1 PUR-1
ANTIOXIDENT RE EVI-1 LF-A1 RB
AP-1 GAG LVF SIE
AP-4 GFI-1 MRE SRE
CCAAT H4TF MTF SRY
CDP HAS+HBS NEUROD1 TFE-3
CEF-1 HBS/XBP NFkB TR
C-MYC HINF NPAS2 TR(DR-4)
COUP-TF HSF PDX-1 TREF-1/2
E47 IKAROS PIT-1 USF-1
EGR XBP-1 XRE
Incubate your sample with the antibody-conjugated beads for 30 minutes
IL-10 TNF-α
1 Add detection antibody and incubate for 30 minutes
2
Add SAPE and incubatefor 30 minutes
3 Detect interactions ona Luminex instrument
4
Procarta Quantitative, Multiplexed Cytokine/Chemokine Assays
Overview
Procarta Cytokine/Chemokine assays use the xMAP technology (mutli-analyte profiling beads) to enable the detection and quantitation of multiple protein targets simultaneously. The Procarta assay kits are compatible with all Luminex and Luminex-based instruments currently available.
Measure Protein and Gene Expression from the Same Sample Well
Panomics’ Procarta Cytokine Assay Kits enable the profiling of up to 33 (check website for an updated list) different cytokines per reaction. When coupled with Panomics’ QuantiGene Plex assays, both gene and protein expression can be
quantitated from the same sample well (supernatant for protein quantitation and cell lysate for mRNA quantitation) enabling the parallel study of gene expression at the RNA and protein level.
Procarta Cytokine Assay Kits
Procarta Cytokine assays simultaneously quantitate cytokines from diverse matrices in 3 hours with a sensitivity of 1 pg/mL/cytokine. Human, Mouse and Rat Cytokine/Chemokine Assay Kits are available in 1- and 10-plate sizes for fixed, off-the-shelf formats, or By Request, customer defined mix-to-order formats. By Request orders are processed and delivered in a pre-mixed ready to use format in ~1 week. Procarta Cytokine Assay Kits contain all the com-ponents required to process cell culture supernatant samples and include: pre-mixed, ready to use, antibody-conjugated
beads; 96-well filter plate and holder; assay and wash buffers; sample buffer; pre-mixed, ready to use detection antibod-ies; Streptavidin-PE (SAPE) fluorescence detection reagent; and premixed antigen standards.
Procarta Standard Diluent Kits
Procarta Cytokine Standard Diluent Kits for plasma and serum, sold separately, contain a single component, species/matrix-specific Standard Buffer, for prepara-tion of antigen standards and dilution of experimental samples (if required). Panomics’ Procarta Cytokine Standard Diluent Kits are designed for use with Procarta Cytokine Assay Kits. Using the appropriate Standard Buffer ensures optimal recovery and sensitivity of the cytokines being analyzed in a serum or plasma matrix.
How It Works
0
5,000
10,000
15,000
20,000
25,000
30,000
35,000
1 10 100 1,000 10,000 100,000
Analyte (pg/ml)
Med
ian
Fluo
resc
ence
Inte
nsity
(MFI
)
IL-1alpha IL-1betaIL-2 IL-3IL-4 IL-5IL-6 IL-10IL-12 (p40) IL-12 (p70)IL-13 IL-17GM-CSF MIP-1alphaEOTAXIN KCRANTES TNFalphaIFNgamma
Standard Curves were generated in cell culture media using Panomics’ 19-plex Procarta Mouse Cytokine Assay Kit. Each analyte has a sensitivity of 10 pg/mL or less and an assay range over 3 logs.
Assay Highlights
Mix and match to create your own plex set from Human, Mouse and Rat analytes from the list below
Complete assay in less than 3 hours
Reagents are supplied at 1X concentration and ready to use
Quantitative measurements from cell culture supernatants, serum or plasma samples
Minimal sample required, only 25 µL
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30,000
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mRN
A E
xpre
ssio
n (M
FI)
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Protein Expression (MFI)
Protein IL-8
mRNA IL-8
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Minutes
mRN
A E
xpre
ssio
n (M
FI)
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350
Protein Expression (MFI)
Protein IL-1β
mRNA IL-1β
Specifications
Sensitivity 1 pg/mL/cytokine
Precision Average Inter-assay CV <10%Average Intra-assay CV <10%
Spike Recovery (Accuracy)
80-120%
Cross Reactivity Negligible
Matrices Cell culture supernatants, serum, plasma
Simultaneous analysis of protein and gene expression from the same sample. Human histocytic lymphoma cells, U-937, were treated with 1 µg/mL of LPS. At various timepoints, cells culture supernatants samples were collected and the corresponding cells were lysed. The supernatants were analyzed for 20 different cytokines using Procarta Human Cytokine Assay. The cell lysates were analyzed for 30 different cytokines using QuantiGene Plex Reagent System. The results of protein and gene analysis of two cytokines, IL-8 and IL-1β, are shown above.
Human 33
L-1-alpha
L-1-beta
IL-2
IL-4
IL-5
IL-6
IL-7
IL-8
IL-10
IL-12(p70)
IL-12(p40)
IL-13
IL-17
ENA-78
EOTAXIN
FGF-basic
G-CSF
GM-CSF
GRO-alpha
iIFN-gamma
IP-10
LEPTIN
MCP-1
MCP-3
MIG
MIP-1-alpha
MIP-2-alpha
NGF
PDGF-BB
RANTES
TGF-beta
TNF-alpha
VEGF
L-1-alpha
IL-1-beta
IL-2
IL-3
IL-4
IL-5
IL-6
IL-10
IL-12(p40)
IL-12(p70)
IL-13
IL-17
IP-10
EOTAXIN
GM-CSF
IFN-gamma
KC
MCP-1
MCP-3
MIP-1-alpha
RANTES
TNF-alpha
VEGF
Mouse 23
IL-1-alpha
IL-1-beta
IL-6
I-CAM
KC
MCP-1
MIP-1-alpha
TNF-alpha
VCAM
Rat 9
The SH2 Domain—a Key to Understanding Phosphotyrosine-Dependent Signal Transduction
SH2 domains are one of the many protein domain families that mediate protein-protein interactions in signal trans-duction. Like other domains, SH2 domains are defined by a conserved region of amino acid residues. The folding characteristics of this sequence of 100-amino acids allow these domains to specifically recognize and bind to phosphotyrosine-containing ligands.
There are approximately 120 different SH2 domains that bind to 110 different proteins in the human genome. These protein-protein interactions involving phosphotyrosines, like those made possible by SH2 domains, are a primary means of recruiting signaling proteins, and thus play a major role in signal transduction.
SH2 domains can be found in enzymes, adaptor proteins, regulatory subunits of signaling proteins, scaffold proteins, transcription factors and oncogenic proteins. These pro-teins are integral to the signaling process because they act as adaptors between receptors and downstream signaling molecules, transmitting signals within cells and regulating the kinase activity of specific proteins.
Protein phosphorylation is a major conduit of information for cellular responses, and defects in SH2 domain-dependent signaling are often directly or indirectly shown to be involved in human diseases.
Phosphotyrosine Profiling Using SH2 Domains
The Procarta SH2 Domain Plex assay is a 30 plex assay capable of profiling identifying differences of measuring SH2 proteins that have bound to phosphorylated tyrosine residues of proteins.
Cos EGF +/- 5ug Cos -, 5ugCos EGF 5'Cos EGF 30Blank
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3BP2
(7)
ABL
2 (1
2)
BTK
(18)
GRA
P (1
9)
CRK
(20)
CRKL
(21)
DA
PP1
(25)
FYN
(26)
GRB
10 (2
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GRB
14 (2
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CSK
(29)
VAV3
(32)
LCK
(34)
LCP2
(35)
MA
TK (3
6)
NSP
1 (3
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GrB
2 (3
8)
P55G
-D1
(41)
P85A
-D1
(42)
P85A
-D2
(43)
P85B
-D1
(44)
P85B
-D2
(45)
PLCG
1-D
1 (4
6)
PTPN
11-D
2 (4
7)
PTPN
6-D
2 (5
1)
SOCS
2 (5
2)
STA
P2 (5
3)
SYK-
D2
(54)
TNS
(55)
SHC1
(56)
P
P
P
P
P
P
Ligand Binding
Receptor Tyrosine Kinase
Receptor phosphorylationand recruitment of SH2 Protein to the RTK
The speci�c SH2 protein binds to the phosphorylated receptorand initiates the cell signaling cascades
SH2-GRB2
SH2-P13K
SH2-PLCY SH2-
Abl
Ligand
Receptor Tyrosine Kinase
Ligand binds to receptor causingphosphorylation of the receptor
SH2-GRB2
SH2-P13K
SH2-PLCY SH2-
Abl
SH2-GRB2
How It Works Assay Highlights
Mix and match to create your own plex set
Complete assay in less than 4 hours
Reagents are supplied at 1X concentration and ready to use
Determine which SH2 domains bind phosphorylated proteins in a given pathway
Key Applications
SH2 Profiling
Peptide affinity screening
Drug binding screening
Phosphoprotein detection using specific antibody
bead1
ABL2
bead1
ABL2
bead1
ABL2
Streptavidin-PE
= Phosphorylated tyrosine proteins
Biotin-conjugatedanti-phosphotyrosine antibody
bead1
bead2
bead3
beadn
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
P
BS-PE
ABL2 CRK LCK GRB10
bead2
CRK
bead3
LCKbead
n
GRB10
Panomics provides Luminex beads conjugated to SH2 proteins.
Treated and untreated cell lysates are prepared containing phosphorylated tyrosine kinases.
SH2 conjugated beads are added to the cell lysates and only the specific SH2 bead will bind to the phosphorylated receptor tyrosine kinases (RTKs).
Anti-phosphotyrosine antibody is added, followed by the addition of the Streptavidin PE. The complex is then analyzed on the Luminex System. The beads that do not have any bound RTKs will have little or no fluorescence.
3BP2 CSK P85B-D1
ABL2 VAV3 P85B-D2
BTK LCK PLCG1-D1
GRAP LCP2 PTPN11-D2
CRK MATK PTPN6-D2
CRKL NSP1 SOCS2
DAPP1 GRB2 STAP2
FYN P55G-D1 SYK-D2
GRB10 P85A-D1 TNS
GRB14 P85A-D2 SHC1
Procarta SH2 Domain Plex
Visit our website to see the most current list.
Create your own SH2 Plex PanelYou have the option to either order
the full 30 plex or choose SH2s from
the panel above to create a panel
from 2-29 plex.
Luminex Technology Overview
Luminex’s xMAP technology is built on proven, existing technology—flow cytometry, microspheres, lasers, digital signal processing and traditional chemistry—that have been combined in a unique way. Featuring a flexible, open-architecture design, xMAP technology can be configured to perform a wide variety of bioassays quickly, cost-effectively and accurately.
Luminex color-codes tiny beads, called microspheres, into 100 distinct sets. Each bead set can be coated with a reagent specific to a particular bioassay, allowing the capture and detection of specific analytes from a sample. Within the Luminex compact analyzer, lasers excite the internal dyes that identify each micro-sphere particle, and also any reporter dye captured during the assay. Many readings are made on each bead set, further validat-ing the results. In this way, xMAP technology allows multiplexing of up to 100 unique assays within a single sample, both rapidly and precisely.
Here’s How It Works
The Luminex System is a flexible analyzer based on the principles of flow cytometry that is designed to meet the needs of any size research laboratory. The system enables you to multiplex (simultaneously measure) up to 100 analytes in a single microplate well, using very small sample volumes. At Panomics though, we offer multiplexed solutions of up to 40 different analytes in a single well. The system delivers fast and cost-effective bioassay results on many assay formats that Panomics offers which include: gene expression, transcription factor profiling, cytokine profiling and SH2 Domain profiling.
The Luminex System is the combination of three core xMAP technologies. The first is xMAP microspheres, a family of 100 fluorescently dyed 5.6 micron-sized polystyrene microspheres that act as both the identifier and the solid surface to build the assay. The second is a flow cytometry-based instrument, the Luminex analyzer, which integrates key xMAP detection components such as lasers, optics, advanced fluidics and high-speed digital signal processors. The third component is the assays that are designed around the microspheres.
xMAP Technology
The xMAP technology uses 5.6 micron polystyrene microspheres which are internally dyed with red and infrared fluorophores. Using different amounts of the two dyes for different batches of micro-spheres, up to 100 different microsphere sets can be created. Each bead is unique with a spectral signature determined by it’s red/infrared dye mixture. The bead is filled with a specific known ratio of the two dyes. As each microsphere carries a unique signature, the xMAP detection system can identify to which set it belongs. Therefore, multiplexing up to 100 tests in a single reaction volume is possible.
Luminex Reader Design
The Luminex reader combines two lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different sets of color-coded polystyrene beads, each bearing a different assay. The Luminex reader is an essential tool that performs the key functions of this multiplex technology:
The bead isimpregnatedwith the dyemixture
5.6 Microns
Bead Set 26
Bead Set 21
10 Unique Infrared Dye Concentrations 10 Unique Red Dye Concentrations
Luminex Performance Highlights
Reduced cost and labor by multiplexing
Shortened time-to-results by favor-able reaction kinetics of liquid bead array approach, with smaller sample requirements
Liquid reaction kinetics give faster, more reproducible results than with solid, planar arrays
Focused, flexible multiplexing in the range of 1 to 100 analytes meets the needs of a wide variety of appli-cations—protein expression profiling, focused gene expression profiling
Fluidics—The reader detects individual beads by flow cytometry. The fluidics system of the reader aligns the beads into single file as they enter a stream of sheath fluid and then enter a flow cell. Once the beads are in single file within the flow cell, each bead is individually interrogated for bead color (analyte) and assay signal strength (PE fluorescence intensity)
Lasers—The reader uses a 532 nm green laser (“assay” laser) is used to excite the PE dye of the assay (Streptavidin-PE). The 635 nm solid state laser (red “classify” laser) is used to excite the dyes inside the beads to determine their “color” or “region” and is also used for doublet discrimination by light scatter
Detectors—The reader has four detectors, one for each of the optical paths shown in the figure below. Detectors are used to measure the fluorescence of the assay, to make bead determination (1-100) and the last to discriminate between single and aggregate beads.
Bead set 12: IL-61835 m� = 500 IU
Assay Detector
532 nmGreen Laser
635 nmRed Laser
Bead Detector 2
Bead Detector 1
DD Detector
U.S. Corporate Headquarters
Panomics, Inc.6519 Dumbarton CircleFremont, CA 94555Toll Free: 877 PANOMICS (1.877.726.6642)Direct: 1.510.818.2600Fax: 1.510.818.2610Email: [email protected] Email: [email protected]: [email protected]
European Headquarters
Panomics SrlVia Sardegna 120060 Vignate-Milano (Italy)Tel: +39.02.95.360.250Fax: +39.02.95.360.992Email: [email protected]: [email protected]: [email protected]
www.panomics.com
6519 Dumbarton CircleFremont, CA 94555Toll Free: 1.877 PANOMICS (1.877.726.6642)T: 510.818.2600F: 510.818.2610www.panomics.com
For pricing and more information visit our website at www.panomics.com or call us at 1.877.726.6642.We also offer custom services. Contact us for details.
© 2007 Panomics, Inc. All rights reserved. Panomics is a trademark of Panomics, Inc. Procarta is a registered trademark of Panomics, Inc. QuantiGene an is a registered trademark exclusively licensed to Panomics, Inc. xMAP and Luminex are registered trademarks of Luminex Corporation. All other trademarks belong to their respective owners. Part #14940
